AbstractPhloem unloading and loading are associated with stem non‐structural carbohydrates (NSCs) accumulation and remobilization in rice (Oryza sativa L.). Four rice recombinant inbred lines (R032, R191, R046, and R146) derived from a cross between Zhenshan 97 and Minghui 63 were used to investigate the contributions of stem large and small vascular bundles (SVBs) to NSCs accumulation and translocation. Before heading, the parenchyma cells in stem cortex tissues (PCs) surrounding SVBs had higher starch density than those surrounding large vascular bundles (LVBs). Moreover, the protein levels of sucrose transporters (SUTs), cell wall invertase, sucrose synthase, and adenosine diphosphate glucose pyrophosphorylase, as well as the phloem plasmodesma densities were higher in SVBs than those in LVBs. After heading, starch density decreased more in PCs surrounding SVBs than in LVBs. Also, the protein levels of SUTs, α‐amylase, sucrose phosphate synthase and sucrose synthase, the phloem plasmodesma densities in SVBs were higher than those in LVBs. The correlations of the number and total cross‐sectional area of SVBs with mass and contribution to yield of transferred NSCs were higher than those of LVBs. Our results suggest that SVBs may have higher contributions to pre‐anthesis stem NSCs accumulation and post‐anthesis translocation than LVBs, which is potentially attributed to the high level of protein and enzyme involved in stem unloading and loading via apoplastic and symplastic pathways.